*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com and include the link in your posting.
*****

These vesicles can also form when cells are dying, maybe they have been in
the same media for too long.

On Mon, Apr 15, 2024, 11:57 AM Kilgore, Jason A. <
[log in to unmask]> wrote:

> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
> https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Alas, I'm sorry to hear that, Michael! Best to just throw the culture out
> and start over. You should check other cell cultures that were in the same
> incubator and give that incubator a thorough cleaning.
>
> Also, there are commercial mycoplasma detection kits you can use to
> confirm. There are fluorescent kits, but they are presumptive and you've
> done that bit already. I'd suggest going with the kits that are PCR-based
> because they are specific and not presumptive (Full disclosure: my company
> sells such kits).
>
> Jason
>
> Jason A. Kilgore
> Supervisor, Cell Analysis Tech Support
> Life Sciences Solutions
> Thermo Fisher Scientific
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Model, Michael
> Sent: Monday, April 15, 2024 11:23 AM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> CAUTION: This email originated from outside of Thermo Fisher Scientific.
> If you believe it to be suspicious, report using the Report Phish button in
> Outlook or send to [log in to unmask]
>
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!DHrFDLindgvbWOQG-7og_69PwmYhJkvAiTDKB_Ti2oPm4GbRurA9Rrobfm-EYW3lhS54JBZ6TFuaxIfXBhaUVB5f$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com__;!!HLrAl2XzZ3iCLg!DHrFDLindgvbWOQG-7og_69PwmYhJkvAiTDKB_Ti2oPm4GbRurA9Rrobfm-EYW3lhS54JBZ6TFuaxIfXBv8gW1uH$
> and include the link in your posting.
> *****
>
> It looks indeed like mycoplasma or something.
> But the first google results gives me "Mycoplasma cells are very small
> bacteria therefore they cannot be detected by visual inspection using a
> visible light microscope". But ours are ~0.5 um and we see them bright and
> clear! In addition, Hoechst staining reveals bright spots. There is not
> always a 100% match with bright-field but we better through away the
> culture.
>
> Thanks again a for good advice!
>
> Mike
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Kilgore, Jason A.
> Sent: Thursday, April 11, 2024 7:57 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!DHrFDLindgvbWOQG-7og_69PwmYhJkvAiTDKB_Ti2oPm4GbRurA9Rrobfm-EYW3lhS54JBZ6TFuaxIfXBhaUVB5f$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com/__;!!HLrAl2XzZ3iCLg!DHrFDLindgvbWOQG-7og_69PwmYhJkvAiTDKB_Ti2oPm4GbRurA9Rrobfm-EYW3lhS54JBZ6TFuaxIfXBugId0v3$
> and include the link in your posting.
> *****
>
> Yes, I believe so, though the signal may be dim compared to the nuclei.
> (using Hoechst 33342 or Hoechst 33258).
>
> Jason
>
> Jason A. Kilgore
> Supervisor, Cell Analysis Tech Support
> Life Sciences Solutions
> Thermo Fisher Scientific
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Model, Michael
> Sent: Thursday, April 11, 2024 4:51 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> CAUTION: This email originated from outside of Thermo Fisher Scientific.
> If you believe it to be suspicious, report using the Report Phish button in
> Outlook or send to [log in to unmask]
>
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!HcgwUADu1Fv7eMBmtMjyyBawgEntpqdcejxqmaYeGtVmgrGzNouPVwCYXgslB0XePijj1E8jfTbQZdGCNcHUxbn0$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com__;!!HLrAl2XzZ3iCLg!HcgwUADu1Fv7eMBmtMjyyBawgEntpqdcejxqmaYeGtVmgrGzNouPVwCYXgslB0XePijj1E8jfTbQZdGCNaMO41X4$
> and include the link in your posting.
> *****
>
> Interesting! If this were so, would Hoechst stain them?
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Kilgore, Jason A.
> Sent: Thursday, April 11, 2024 7:44 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!HcgwUADu1Fv7eMBmtMjyyBawgEntpqdcejxqmaYeGtVmgrGzNouPVwCYXgslB0XePijj1E8jfTbQZdGCNcHUxbn0$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com/__;!!HLrAl2XzZ3iCLg!HcgwUADu1Fv7eMBmtMjyyBawgEntpqdcejxqmaYeGtVmgrGzNouPVwCYXgslB0XePijj1E8jfTbQZdGCNQCbALOp$
> and include the link in your posting.
> *****
>
> Michael,
>
> I hope I'm wrong about this, but it reminds me of images of
> mycoplasma-infected cells. Those cells also don't look very healthy. Could
> they be infected?
>
> Jason
>
> Jason A. Kilgore
> Supervisor, Cell Analysis Tech Support
> Life Sciences Solutions
> Thermo Fisher Scientific
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Model, Michael
> Sent: Thursday, April 11, 2024 4:38 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> CAUTION: This email originated from outside of Thermo Fisher Scientific.
> If you believe it to be suspicious, report using the Report Phish button in
> Outlook or send to [log in to unmask]
>
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFqmJrTAD$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFuWcJGpZ$
> and include the link in your posting.
> *****
>
>
> Thank you, Tim,
> But I don't think this is it. I placed one of the images here:
>
>
> https://urldefense.com/v3/__https://www.dropbox.com/scl/fo/3p8yfmm5rqoo1rqj36bkd/AJOTl2uNKOh6DSXztwO2HWI?rlkey=h771q3h1abhtssv327dng2yre&dl=0__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFgMjkrAq$
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Tim F
> Sent: Thursday, April 11, 2024 5:24 PM
> To: [log in to unmask]
> Subject: EXT: Re: vesicles
>
> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
>
> https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFqmJrTAD$
> Post images on
> https://urldefense.com/v3/__http://www.imgur.com/__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFohxnDzh$
> and include the link in your posting.
> *****
>
> Hi Michael, just to confirm, if your experience is similar to the video
> below, do you mean the dark spots? Thanks to Nikon for the vid!
>
>
> https://urldefense.com/v3/__https://youtu.be/Rb-Q3wLsbIk?si=f8GSrHrkTEYmZg1t__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFk1AOE-7$
>
> I remember thinking those are protein mis-folding aggregates. I don't
> think you would see endosomes with that kind of contrast, and vesicles are
> too small at 60nm to see with transmitted light.
>
> Best,
>
>
> T
>
> Timothy Feinstein, Ph.D.
>
> On Thu, Apr 11, 2024 at 3:53 PM Model, Michael <[log in to unmask]> wrote:
>
> > *****
> > To join or leave the confocal microscopy listserv or to change your
> > email address, go to:
> > https://urldefense.com/v3/__https://urld/__;!!HLrAl2XzZ3iCLg!DHrFDLind
> > gvbWOQG-7og_69PwmYhJkvAiTDKB_Ti2oPm4GbRurA9Rrobfm-EYW3lhS54JBZ6TFuaxIf
> > XBu5CQ0N7$
> > efense.com%2Fv3%2F__https%3A%2F%2Furld%2F__%3B!!HLrAl2XzZ3iCLg!HcgwUAD
> > u1&data=05%7C02%7Cmmodel%40KENT.EDU%7C18f4d987cc58409d88f908dc5a83321e
> > %7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C638484766728876524%7CUnk
> > nown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWw
> > iLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=dMXTIHvNfI6nTPY2rjLCXzxAorWnboIKY5c
> > 4KgMnLQs%3D&reserved=0
> > Fv7eMBmtMjyyBawgEntpqdcejxqmaYeGtVmgrGzNouPVwCYXgslB0XePijj1E8jfTbQZdG
> > CNWi6jkSt$
> > efense.com%2Fv3%2F__https%3A%2F%2Flist%2F__%3B!!HLrAl2XzZ3iCLg!B9GBDpQ
> > nx&data=05%7C02%7Cmmodel%40KENT.EDU%7Cedbc032c0120404b608a08dc5a815205
> > %7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C638484758673745277%7CUnk
> > nown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWw
> > iLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=e6%2Fjtgn4Xq4zGhPwdOJ7CiaDaEUdiUzIN
> > p5m2A0psiw%3D&reserved=0
> > u1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_ukn
> > NFn86y8bg$
> > s.umn.edu%2Fcgi-bin%2Fwa%3FSUBED1%3Dconfocalmicroscopy%26A%3D1&data=05
> > %7C02%7Cmmodel%40KENT.EDU%7C560e6be681534059186e08dc5a6e4300%7Ce5a06f4
> > a1ec44d018f73e7dd15f26134%7C1%7C0%7C638484676935174256%7CUnknown%7CTWF
> > pbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6M
> > n0%3D%7C60000%7C%7C%7C&sdata=6AJ4ERdzHUxtlMrfZG2aSTCg8fzFtpz9J3jiAK37H
> > i0%3D&reserved=0 Post images on
> >
> https://urldefense.com/v3/__http://www.imgur.com/__;!!HLrAl2XzZ3iCLg!B9GBDpQnxu1Zw-80F71B9xaYdV0k6k1-N7cd7igG9A0s7AAXrs4NrHC1Yq7PitL8kMdg_Gd8sKH_uknNFohxnDzh$
> and include the link in your posting.
> > *****
> >
> > Dear List,
> >
> > If you look at cultured cells, such as HeLa, under bright field, you
> > clearly see many vesicles about 1 um. Does anyone know what they are?
> > (They are not acidic). The stupidest thing is that I once found a
> > paper that investigates them, but now I cannot find that paper!
> >
> > Mike
> >
> CAUTION: EXTERNAL SENDER Do not click any links, open any attachments, or
> REPLY to the message unless you trust the sender and know the content is
> safe.
>